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PMID: 6312834 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels.

Analytical biochemistry ·Vol. 132 ·No. 1 ·1983-07-01 ·Pages 14-9

Tautz D, Renz M

Abstract

A procedure for quick and simple elution of DNA from agarose gels is presented. After electrophoresis, bands of interest are cut out of the gel and the slices are equilibrated in a neutral salt buffer. The slices are then frozen and centrifuged through a filtration assembly whereby the DNA-containing buffer is squeezed out. The method is simple, quick, and suitable for the safe handling of small amounts of DNA (less than 1 microgram). The isolated DNA is susceptible to any enzymatic reaction and also to chemical sequencing. The method is most useful for rapid preparation of specifically end-labeled DNA fragments (e.g., for sequencing), but may also be utilized for any other preparative applications.

MeSH Terms
Bacteriophage lambda/metabolism DNA/isolation & purification,metabolism DNA Restriction Enzymes Electrophoresis, Agar Gel Hydrogen-Ion Concentration Molecular Weight Salts Sepharose
Chemicals
Salts DNA Sepharose DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tautz D
Renz M
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
0003-2697
Published
1983-07-01
Pages
14-9
Language
English
Region
United States
NLM ID
0370535
Subset
IM
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